It depends on your study plan, what your are looking for. For eg. you can perform sperm function tests, DNA damage test (comet assay, SCSA etc), extract DNA (mutation and polymorphism studies), RNA (expression pattern of genes) and proteins (proteomic profile using LC-MS or specific protein using western blot) (note: for this you need to separate sperm cell using DGC or swim up, free from contaminants) etc. You  have to specify what exactly is your study plan with those sperm pellet!! 

Hope this is of help.

Thanks

You can use them for structural (morphology, DNA integrity, damage in mid piece or tail etc) , functional (hyper activation, Acrosome reaction, cervical mucous penetration etc) or biochemical assays (acrosin activity, ROS etc) depending on your objectives.

I agree with the colleagues that wrote previously. The firsat question is "wath do you want to do?" so, you can search if using the sperm pellet is the best to your research.

We use sperm pellets to conduct high resolution light microscopy and transmission electron microscopy after embedding in  appropriate resins. These studies are particularly useful to review sperm pathologies at high magnification and the involvement of different sperm organelles in sperm pathologies of the head, mid piece and flagellum in patients with extreme asthenozoospermia, sperm immotility and/or severe teratozoospermia.

Than you very much all of you,

i want to measure ROS, cervical mucus penetration, DNA integrity and mitochondrial activity.

Well, if that are your idea, dependong on the species, some of the tests that you will run up are better to do with resuspended spems with special diluent for each different measurement. There are several tests developed for those measurements, that are more or less similar but with special issues by species. I recommend analyse DNA integrity withouth pelleting the semen.

Thank you, Humberto Sir,

Can you provide some tests  with referances which can i performed?

It depends on wath equipement you have access. You can write me and I´ll send you some tips in order of the equipment you have.

SIr, I want to study DNA in bovine sperm through Acridine-Orange stain through fluorescent microscope.

And mitochondrial activity, about which i have no idea. 

Requiring guidance in this regard.

this protocol am using for my study and its too easy and time consuming method.

Sperm DNA integrity was evaluated using the acridine orange

method described by Varela Junior et al. (2012), where the

metachromatic colorant acridine orange emits green fluorescence

in reaction to double-stranded DNA and an orange or red fluorescence

in reaction to single-stranded DNA, identifying a break in the

DNA. The sperm sample (45 L) was diluted in 50 L TNE (0.01 M

Tris–HCl; 0.15 M NaCl; 0.001 M EDTA; pH 7.2). After 30 s, 200 L

of Triton solution 1× were added, and 30 s later, 50 L of acridine

orange were added(2 mg/mL indeionizedH2O). The evaluation was

conducted after 5 min without exceeding 1 min of slide exposure.

The sperm presenting with green fluorescence contained intact

DNA, and those presenting with red or orange fluorescence contained

denatured DNA. The rate of DNA integrity was determined

by the proportion of sperm emitting green fluorescence compared

with the total number of sperm analyzed (green, red or orange).

Can you give me the full length article of Varela Junior et al. (2012) ?

Herewith i attached as you requested paper please find it

thank you

thank you